A Novel Regulatory Pathway for K+ Uptake in the Legume Symbiont Azorhizobium caulinodans in Which TrkJ Represses the kdpFABC Operon at High Extracellular K+ Concentrations

ABSTRACT Bacteria have multiple K+ uptake systems. Escherichia coli, for example, has three types of K+ uptake systems, which include the low-K+-inducible KdpFABC system and two constitutive systems, Trk (TrkAG and TrkAH) and Kup. Azorhizobium caulinodans ORS571, a rhizobium that forms nitrogen-fixing nodules on the stems and roots of Sesbania rostrata, also has three types of K+ uptake systems. Through phylogenetic analysis, we found that A. caulinodans has two genes homologous to trkG and trkH, designated trkI and trkJ. We also found that trkI is adjacent to trkA in the genome and these two genes are transcribed as an operon; however, trkJ is present at a distinct locus. Our results demonstrated that trkAI, trkJ, and kup were expressed in the wild-type stem nodules, whereas kdpFABC was not. Interestingly, Δkup and Δkup ΔkdpA mutants formed Fix– nodules, while the Δkup ΔtrkA ΔtrkI ΔtrkJ mutant formed Fix+ nodules, suggesting that with the additional deletion of Trk system genes in the Δkup mutant, Fix+ nodule phenotypes were recovered. kdpFABC of the Δkup ΔtrkJ mutant was expressed in stem nodules, but not in the free-living state, under high-K+ conditions. However, kdpFABC of the Δkup ΔtrkA ΔtrkI ΔtrkJ mutant was highly expressed even under high-K+ conditions. The cytoplasmic K+ levels in the Δkup ΔtrkA ΔtrkI mutant, which did not express kdpFABC under high-K+ conditions, were markedly lower than those in the Δkup ΔtrkA ΔtrkI ΔtrkJ mutant. Taking all these results into consideration, we propose that TrkJ is involved in the repression of kdpFABC in response to high external K+ concentrations and that the TrkAI system is unable to function in stem nodules. IMPORTANCE K+ is a major cytoplasmic cation in prokaryotic and eukaryotic cells. Bacteria have multiple K+ uptake systems to control the cytoplasmic K+ levels. In many bacteria, the K+ uptake system KdpFABC is expressed under low-K+ conditions. For years, many researchers have argued over how bacteria sense K+ concentrations. Although KdpD of Escherichia coli is known to sense both cytoplasmic and extracellular K+ concentrations, the detailed mechanism of K+ sensing is still unclear. In this study, we propose that the transmembrane TrkJ protein of Azorhizobium caulinodans acts as a sensor for the extracellular K+ concentration and that high extracellular K+ concentrations repress the expression of KdpFABC via TrkJ.

On the origins and evolution of trans-splicing of bursicon in mosquitos

Broad transcriptomic sequencing of eukaryotes has revealed the ubiquity of splicing of nuclear genes. While the vast majority of splicing events join segments of the same RNA transcript, various studies have found a few intriguing cases of trans-splicing of introns, in which splicing events within protein coding regions join segments of different RNA transcripts. The most structurally intricate case known involves the bursicon gene in mosquitos, in which an internal exon is encoded at a distinct locus, requiring multiple trans-splicing events form the mature mRNA. This arrangement is known to be ancestral to mosquitos, however the exact timing of the origin of trans-splicing and the history of the bursicon gene within mosquitos is unknown. Taking advantage of the recent availability of genomes from various Anopheles mosquitos and from relatives of mosquitos, I determined trans versus cis encoding of bursicon across Culicomorpha. I conclude that trans-splicing emerged in the last common ancestor of mosquitos, and that trans-splicing has been retained in all 19 studied Anopheles species. The retention of trans-splicing could indicate functional importance of this arrangement, or could alternatively reflect the rarity of mutations giving rise to viable allelic alternatives.

Smoking-Specific Locus of Control

A smoking-specific locus of control scale was developed by modifying a drinking-related locus of control scale. The new scale, Rotter's I-E scale, and a smoking questionnaire were administered to 70 subjects. Estimates of reliability indicated that the new scale had reasonably high internal consistency. Scores differed as a function of subjects' reports of their addiction to cigarettes but not as a function of their current smoking status. 15 addicted individuals scored more externally on the new scale than did 24 nonaddicted individuals. There were no differences for scores on the Rotter I-E scale as a function of reported addiction or smoking status. The new scale appears to be a more sensitive measure of locus of control than Rotter's scale. The two locus of control scales were not significantly correlated, indicating their independence. Individuals apparently maintain a distinct locus of control for smoking rather than having a general expectation for control.

On the origin of meiotic reproduction: a genetic modifier model.

We study the conditions under which a rare allele that modifies the relative rates of meiotic reproduction and apomixis increases in a population in which meiotic reproduction entails selfing as well as random outcrossing. A distinct locus, at which mutation maintains alleles that are lethal in homozygous form, determines viability. We find that low viability of carriers of the lethal alleles, high rates of selfing, dominance of the introduced modifier allele, and lower rates of recombination promote the evolution of meiosis. Meiotic reproduction can evolve even in the absence of linkage between the modifier and the viability locus. The adaptive value of meiotic reproduction depends on the relative viabilities of offspring derived by meiosis and by apomixis, and on associations between the modifier and the viability locus. Meiotic reproduction, particularly under selfing, generates more diverse offspring, including those with very high and very low viability. Elimination of offspring with low viability generates positive associations between enhancers of meiotic reproduction and high viability. In addition, partial selfing generates positive associations in heterozygosity (identity disequilibrium) between the modifier and the viability locus, even in the absence of linkage. The two kinds of associations together can compensate for initial reductions in mean offspring viability under meiotic reproduction.